1. IL-9–expressing TH cells are highly enriched in CCR4+/CCR8+ effector memory TH cells.
IL-9–expressing THcells are highly enriched in CCR4+/CCR8+effector memory THcells. (A) Naïve, central memory (TCM), and effector memory (TEM) TH cells were sorted from PBMCs of healthy donors according to the expression of CCR7, CD45RA, and CD25 and activated with αCD3/CD2/CD28 beads. The production of IL-9 and IFN-γ was assessed by flow cytometry at the indicated time points. (B) Effector memory TH cells were repeatedly activated with αCD3/2/28 beads, and IL-9 production was measured at the indicated time points by flow cytometry. (C) CD4+/CD25− T cells were sorted according to the expression of the indicated chemokine receptors into positive (black bars) and negative (white bars) populations and activated with αCD3/CD2/CD28 beads. On day 2, production of IFN-γ, IL-13, IL-17, and IL-9 was simultaneously analyzed by flow cytometry (IL-9single+ = IL-9+/IFN-γ−/IL-13−/IL-17− TH cells). ns, not significant. (D) Representative plots of CXCR3-, CCR4-, CCR8-, and CCR6-positive and negative memory T cells, stimulated as in (C). (E) CCR4 and CCR8 expression in freshly isolated CXCR3−/CCR6−/CD25− memory TH cells. (F to H) CCR4+/CCR8− and CCR4+/CCR8+ CD25− memory TH cells were sorted from healthy donors and activated with αCD3/CD2/CD28 beads. Production of cytokines was assessed on day 2 (F) or day 6 (G) as in (C). (H) Representative example of cytokine expression in CCR4+/CCR8+ CD25− memory TH cells before and at different time points after activation. Data are representative of independent experiments with at least four donors (B, G, and H) or at least six donors (A and C to F) and presented as means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < Statistics: One-way ANOVA, followed by Tukey multiple comparison test (A), or paired two-tailed t test (C, F, and G).
Claire Micossé et al. Sci. Immunol. 2019;4:eaat5943
Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works